scrutinizing barremian coprolite inclusions to …€¦ · lites are among the most abundant...

Annales Societatis Geologorum Poloniae (2018), vol. 88: 203 – 221 doi: https://doi.org/10.14241/asgp.2018.014

Las Hoyas is an upper-Barremian continental Kon-servat-Lagerstätte located in Cuenca, central Spain, that has yielded a vast diversity of plant and animal body fossils and ichnofossils (Buscalioni and Poyato-Ariza, 2016). Copro-lites are among the most abundant remains, with almost 2,000 coprolites collected. A total of twelve different coprolite morphotypes have been described: spiral, circular, irregular, elongated, rosary, ellipsoidal, cylinder, bump-headed lace, fir-tree, cone, straight lace, and thin lace coprolites (Barri-os-de Pedro et al., 2018a). The Las Hoyas ichnoassemblage contains not only an exceptional diversity of coprolite mor-phologies, but also provides the opportunity to explore the taphonomy of inclusions and infer the digestive strategies of the coprolite producers. Most of the rich Mesozoic coprolite assemblages are from marine and transitional environments (e.g., Eriksson et al., 2011; Niedźwiedzki et al., 2016, Luo et al., 2017), while the Las Hoyas coprolite association was produced in a lacustrine carbonate environment, interpret-ed as a stationary, subtropical, inland wetland, dominated by periphyton and microbial mats (Fregenal-Martínez and Meléndez, 2016; Fregenal-Martínez et al., 2017). There-fore, bacterial sealing has been proposed as one of the most influential processes behind the exceptional preservation at

Las Hoyas (Buscalioni and Fregenal-Martínez, 2010; Inie-sto et al., 2013, 2016; Guerrero et al., 2016), contributing decisively to the protection of the coprolite inclusions.

In a previous contribution, a dichotomous key to delimit the Las Hoyas coprolite morphotypes was elaborated, indi-cating that these coprolites mostly correspond to carnivorous producers with ichthyophagous diets, such as crocodiles, urodelans and different kind of fishes (Barrios-de Pedro et al., 2018a). The aim of this paper is to describe and deter-mine the taphonomic properties of the inclusions contained in the Las Hoyas coprolites; these properties were studied for each morphotype. The combination of such properties in every morphotype is discussed to differentiate the patterns of digestive processes and infer some of the digestive strat-egies of the producers. In general, this study aims to explore the patterns of digestive strategies to extrapolate which was the most dominant in this Barremian ecosystem. The diges-tive strategies are reflected in the abundance of undigested food and the lines of evidence left by digestive processes, such as fermentation and enzymatic degradation (see Ste-vens and Hume, 1995; Furness et al., 2015). Because diges-tive strategies can be shared by different groups (e.g., fish and amphibians), the present authors here evaluate the cor-

SCRUTINIZING BARREMIAN COPROLITE INCLUSIONS TO RECORD DIGESTIVE STRATEGIES

Sandra BARRIOS-DE PEDRO & Ángela D. BUSCALIONI

Departamento de Biología (Paleontología) and Centro para la Integración en Paleobiología (CIPb), Universidad Autónoma de Madrid, Cantoblanco 28049 Madrid, Spain;

e-mails: [email protected]; [email protected]

Barrios-de Pedro, S. & Buscalioni, D. Á., 2018. Scrutinizing Barremian coprolite inclusions to record digestive strategies, Annales Societatis Geologorum Poloniae, 88: 20 – 221.

Abstract: The exceptional preservation of the Las Hoyas coprolites allows the taphonomic study of inclusions on twelve morphotypes and twenty-three specimens. Non-destructive techniques were applied to study the digestion features (pitting, corrosion lines, shape of the fractures at the ends) and the arrangement, number, and size of in-clusions. An analysis based on non-metric, multidimensional scaling ordination identified the similarities among the inclusion features and morphotypes. The morphotypes are clustered on the basis of the way of ingestion and the digestive process. The authors recognize three digestive strategies for the Las Hoyas coprolites: (1) ingestion of prey with limited processing in the mouth, scarce to less effective acid secretions, and/or defecation in a short period of time; (2) ingestion of the prey with mastication prior to deglutition, and defecation over a longer period of time; (3) mastication and long retention time of food in the digestive system with more effective acid secretions. This study is a first step in the understanding of the feeding ecology of the Las Hoyas Barremian lentic ecosystem, based on coprolites.

Key words: Cretaceous, Las Hoyas, lacustrine, taphonomy, digestive process, feeding ecology.

Manuscript received 19 April 2018, accepted 19 October 2018

INTRODUCTION

204 Sandra BarrIOS-dE PEdrO & ÁngEla d. BUSCalIOnI

relation between morphotypes and inclusions, rather than morphotypes and producers (Hunt et al., 2012). This study is the first step towards understanding the feeding ecology of the Las Hoyas continental Barremian wetland.

GEOLOGICAL SETTINGThe Las Hoyas fossil site is a Konservat-Lagerstätte in

the La Huérguina Formation (upper Barremian). It is locat-ed in the Serranía de Cuenca (east-central Spain; Fig. 1), which is part of the Iberian Range. This locality consists of finely laminated limestones, composed almost entirely of calcium carbonate (Fregenal-Martínez and Meléndez, 2016; Fregenal-Martínez et al., 2017). The currently accepted depositional model for the laminated facies of Las Hoyas distinguishes two groups of facies associations (Buscalioni and Fregenal-Martínez, 2010). One association type would have been deposited during wetter periods by traction or suspension fallout of allochthonous carbonate, while the other type would have been formed by the autochthonous production of carbonate, linked to the growth of microbial mats during drier periods of a low water column. The alter-nation of both associations is related to a seasonal, subtrop-ical habitat with a regional climate, alternating between wet and dry seasons (Fregenal-Martínez and Meléndez, 2016). The depositional system corresponds to a continental sedi-mentary environment, in which Las Hoyas would have been a shallow lake, being part of an inland freshwater system of wetlands on a regional scale (Poyato-Ariza et al., 1998; Bailleul et al., 2011, Fregenal-Martínez et al., 2017). This scenario explains the coexistence of terrestrial and aquatic organisms and traces.

The Las Hoyas ichnoassemblage

The ichnoassemblage is characterized by the aquatic Mermia ichnofacies, which at Las Hoyas shows limited di-versity of invertebrate aquatic traces. The locality also con-tains vertebrate terrestrial footprints (Buatois et al., 2000; Gibert et al., 2016). The aquatic trace fossils consist of very shallow-tiered, endogenic, invertebrate burrows and trails, restricted to some bedding planes. The deposit exhib-its minor effects of bioturbation, favouring the exceptional preservation of the finely laminated, primary sedimentary fabric. Therefore, the poor and shallow Mermia ichnofa-cies confirms that the substrate was highly stressed by the limited availability of oxygen and anoxia would have been a key factor in promoting the exquisite preservation of fos-sils, together with bacterial sealing (Buscalioni and Frege-nal-Martínez 2010).

METHODS AND MATERIALSMaterials

The coprolites studied are housed at the Museo de las Ciencias de Castilla-La Mancha (MCCM) and belong to the Las Hoyas (LH) collection. A summary of the main charac-teristics for the twelve morphotypes described in Barrios-de Pedro et al. (2018a) is provided in Table 1. Every morpho-type was subsumed into the morphological classification of faeces, which also includes the producers, provided by Hunt and Lucas (2012, see Table 1).

Most of the Las Hoyas coprolites are preserved in part and counterpart. Each slab is identified by the letters ‘a’ or ‘b’ af-ter the specimen number. Specimen numbers not followed by letters correspond to unsplit specimens (i.e., specimens that are whole). Mostly, coprolites expose a section charac-terized by two linear dimensions (length and diameter, see Table 2). The surface of these sections is irregularly shaped, and different levels of coprolite matrix are exposed. The diameter ranges were categorized in the following states: 1 = ≤5 mm; 2 = 5–9.9 mm; 3 = 10–19.9 mm; 4 = ≥20 mm.

Techniques

In order to explore the inclusions in the coprolite matrix, non-destructive techniques were employed. Macro-scale photos of coprolites were taken, using a D5100 Nikon Reflex camera at the Biology Department of Universidad Autónoma de Madrid (UAM). Other techniques, such as Scanning Electron Microscope (SEM) analyses with a low- vacuum Environmental Scanning Electron Microscope (ESEM_QUANTA200) and a 3D high-resolution micro-scope with interferometric and profilometry (Leica DCM8) were carried out at the Museo Nacional de Ciencias Natu-rales (MNCN) in Madrid. The ImageJ software (Fiji) was used for measurements (Rasband, 1997–2018).

Criteria and features

A set of features of the inclusions were obtained to infer the digestive strategies characterizing the Las Hoyas co-

Fig. 1. Location maps. A. Simplified map of the Iberian Penin-sula showing the location of Cuenca. B. Simplified geological map of the Las Hoyas area.

205SCRUTINIZING BARREMIAN COPROLITE INCLUSIONS TO RECORD DIGESTIVE STRATEGIES

prolite association. The digestive strategies are discussed following the criteria used for the ‘gut-reactor’ digestion model (Penry and Jumars, 1987; Stevens and Hume, 1995; Furness et al., 2015). The gut-reactor model combines the gut architecture, gut capacity, intake level, dietary compo-sition, digesta passage rate, and digestion rate (Penry and Jumars, 1987). Therefore, in order to integrate such parame-ters to discuss the digestive strategy, the following variables were explored in a comprehensive manner (see Table 2): (1) the abundance of undigested food, in order to estimate the intake level; (2) the identification of inclusions, in order to evaluate the dietary composition; (3) the size of inclu-sions and (4) the arrangement of inclusions in the coprolite matrix, in order to discuss digesta and digestion rates; and finally (5) some features related to the digestive process, in order to obtain some values related to the chemical process (Fernández-Jalvo and Andrews, 2016).1) Number of inclusions. The counting of remains was per-

formed from photographs at the macroscale and the vis-ible inclusions in every coprolite were categorized into four states (1, N<15; 2, N = 15–30; 3, N = 31–45 and 4, N >45). The smallest remains would correspond on average to 0.2 mm; those observed with SEM analyses were not taken into account for these data.

2) Identification of inclusions. When possible, inclusions were identified considering their nature, their anatomi-cal elements and/or their taxonomy. An initial identifi-cation was done at the macroscale with a magnifier and SEM then was used to determine the external structures.

3) Size of inclusions. The total length of the coprolites, their maximum diameter, and data on the size of their inclusions are given in millimetres (mm). The scale bars of the SEM analyses are provided in microns (µm). If the number of inclusions in a coprolite was less than 15, all of the inclusions were measured. When the number of inclusions exceeded 15, measurements of all differ-ent inclusion sizes were taken (from the smallest to the largest). In both cases, the minimum and maximum val-ues of the length of the inclusions of each coprolite are given.

4) The arrangements of inclusions in the coprolite matrix are described as fragmented (groups of remains with no anatomic relationship with each other), articulated (some of the remains in anatomical articulation), and isolated (ungrouped and widely separated remains).

5) Action of digestive process. As the main feature of di-gestion processes is the etching of bone surfaces by stomach acids (Fernández-Jalvo and Andrews, 2016), it is possible to record the pitting and corrosion lines on the surfaces of coprolite inclusions; however, the chemical changes caused by digestive enzymes are dif-ficult to observe (Fernández-Jalvo and Andrews, 2016). The following features were studied by SEM analyses: pitting on inclusions and corrosion lines. The variable named ‘chemical signals’, with three stages, is consid-ered to summarize the combination of pitting and corro-sion: light (slight pitting and no/slight corrosion lines); moderate (noticeable pitting and some corrosion lines, or slight pitting and profuse corrosion lines); and heavy (great pitting and marked corrosion lines). In addition,

the shape of fractures at the ends of the inclusions was indicated, if exposed (i.e., blunt, pinched, straight or ir-regular); when two or more of them were present, this variable was codified as ‘all types’ (Table 2).

Morphotypes ordination

The ordering of the morphotypes was explored on the basis of a distance or dissimilarity matrix (Bray-Curtis dis-similarity index, PAST ver. 2.17; Hammer et al., 2001), using the features of undigested food and patterns of the digestive chemical processes per specimen (see Appen-dix 1). Non-metric, multidimensional scaling (NMDS) is an indirect gradient analysis approach, which produces an ordination and attempts to represent the pairwise dissimi-larity between objects in a low-dimensional space. NMDS allows the testing of the relationships between the inclusion properties and the digestive strategies proposed for the mor-photypes. The best-fit solution of NMDS was assessed by the stress of the ordination, which measures how reliably the multidimensional relationships among the samples are represented in the two-dimensional ordination plot. Useful two-dimensional plots should have a stress value smaller than 0.2 (Zuschin et al., 2006).

RESULTSStudy of properties of inclusions

The study of the properties of the inclusions was carried out on twenty-three specimens that belong to the morpho-types previously indicated. Figures 2 and 3 show exam-ples of coprolites (macroscale photos) and their inclusions (SEM photos). Table 2 includes all the described features explained in the Material and Methods section. These ta-bles comprise the coprolite linear dimensions (length and diameter), a description of the inclusions of the specimen, and the resulting patterns of the digestive chemical pro-cesses. See Appendix 2 for a detailed description of the results.

Size of inclusions by morphotype

Generally, the average of the size of inclusions for the Las Hoyas coprolites is less than 2 mm (Fig. 4). Circular, bump-headed lace, fir-tree, rosary, straight lace, and thin lace morphotypes show less variation in the size of their inclusions. The median value is asymmetric for cylinder, cone, ellipsoidal and elongated morphotypes, indicating an unequal variation of inclusions size, significant among the large inclusions and minor among the smaller inclusions. The one specimen corresponding to the spiral morphotype is not representative in this sample, because it only shows two exposed inclusions. The irregular morphotype contains a great number of outliers increasing the mean value up to 3 mm in size. An increase in the diameter of the faecal mass is related to longer inclusions (Fig. 5), which is evident in irregular, elongated, and cone specimens and one of the cyl-inder specimens of more than 7 mm in diameter.


Bum

p-he

aded

lace

Ros

ary

Stra

ight

lace

Thi

n la

ceE

long

ated

Cyl

inde

r

One

of t

he e

nds w

ith a

big

bu

lge

(bb)

, at l

east

twic

e as

w

ide

as th

e re

st o

f the

cop

ro-

lite.

The

ove

rall

shap

e is

elo

ngat

ed c

ord.

Serie

s of w

ide

segm

ents

with

bu

mps

sepa

rate

d by

con

stric

-tio

ns (c

). Th

e ov

eral

l sha

pe is

el

onga

ted

with

bum

ps.

Long

and

stra

ight

, unf

olde

d,

with

a ro

ughl

y si

mila

r dia

me-

ter t

hrou

ghou

t the

leng

th.

The

over

all s

hape

is e

long

ated

co

rd.

Fold

ed o

nto

them

selv

es, w

ith

a ro

ughl

y id

entic

al d

iam

eter

th

roug

hout

thei

r len

gth.

The

ov

eral

l sha

pe is

ribb

on-li

ke.

The

over

all s

hape

is re

ctan

gu-

lar,

at le

ast t

hree

tim

es lo

nger

th

an w

ide.

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t end

s.

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over

all s

hape

is st

rong

ly

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gate

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ith a

roug

hly

con-

stan

t wid

th.

Rou

nded

end

s.

bbc

Mor

pho:

NE

Subm

o: N

EPr

oduc

er: N

E

Mor

pho:

NE

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er: N

E

Mor

pho:

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n Li

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pho:

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ngat

ed

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ucer

:Fi

sh, A

nura

, Che

loni

i, C

roco

-dy

lia, T

hero

poda

Tabl

e 1

The

Las H

oyas

cop

rolit

e as

soci

atio

n (u

pper

Bar

rem

ian,

Cue

nca,

Spa

in).

Ever

y m

orph

otyp

e in

clud

es

a de

scrip

tion

and

a dr

awin

g. T

he lo

wer

line

cor

resp

onds

to th

e cl

assi

ficat

ion

cate

gorie

s use

d in

Hun

t and

Luc

as (2

012)

, an

d co

mpr

ises

: Mor

pho–

mor

phot

ype;

Sub

mo

– su

bmor

phot

ype;

Pro

duce

r – c

opro

lite

prod

ucer

. A

bbre

viat

ions

use

d fo

r mor

phot

ypes

are

: A –

cyl

indr

ical

, elo

ngat

ed; B

– c

ylin

dric

al, s

hort;

C

– su

brou

nded

; F –

spira

l; H

– th

in, o

void

; I –

thin

, lin

ear;

K –

irre

gula

r sha

pe; N

E –

no e

quiv

alen

ce


Con

eFi

r-tr

eeC

ircu

lar

Elli

psoi

dal

Irre

gula

rSp

iral

Dia

met

er in

crea

ses t

hrou

ghou

t th

e lo

ngitu

dina

l axi

s, no

con

-st

rictio

ns. M

axim

um d

iam

eter

oc

curs

at o

ne e

nd. T

he o

vera

ll sh

ape

is c

one

to te

ar-d

rop.

Sequ

ence

of ‘

bum

ps’ t

hat

decr

ease

pro

gres

sive

ly fr

om

a w

ide

end

to a

ver

y na

rrow

en

d. T

he o

vera

ll sh

ape

is tr

iang

ular

.

Rat

her fl

at, b

ut n

ot a

s flat

as

a di

sc. T

he o

vera

ll sh

ape

is

a su

brou

nded

and

flat

tene

d sp

hero

id.

Dia

met

er w

ider

at t

he m

iddl

e,

and

maj

or a

xis t

wic

e or

thre

e tim

es lo

nger

than

min

or a

xis.

The

over

all s

hape

is ro

ughl

y ov

oid.

Not

gro

uped

as d

efine

d m

or-

phot

ype.

With

a v

arie

d m

osai

c of

shap

es.

With

spira

l mar

ks. S

trips

with

re

gula

r wid

ths.

The

over

all

shap

e is

elo

ngat

ed.

Mor

pho:

BSu

bmo:

B2

cylin

dric

al sh

ort

Prod

ucer

: ?Fi

sh

Mor

pho:

CSu

bmo:

NE

Prod

ucer

: NE

Mor

pho:

CSu

bmo:

NE

Prod

ucer

: NE

Mor

pho:

HSu

bmo:

Thi

n-ov

oid

Prod

ucer

: Fis

h

Mor

pho:

KSu

bmo:

Irre

gula

rPr

oduc

er: F

ish

Mor

pho:

FSu

bmo:

F2

Am

phip

olar

Spi

ral

Prod

ucer

: Fis

h


Fig. 2. SEM and BSE images of coprolite specimens from the Las Hoyas locality (upper Barremian, Cuenca, Spain). A. Bump-headed lace coprolite, MCCM-LH15993a. B. MCCM-LH15993a, hollow vertebra. C. MCCM-LH15993a, plant remains with parallel fibres. D. MCCM-LH15993a, vegetal epidermis. E. Cylinder coprolite, MCCM-LH9475A. F. MCCM-LH9475A, sequence of teeth or the internal mould of a tiny, grooved, undetermined fragment (? Mollusc, gastropod) indicated by arrow. G. Cylinder coprolite, MCCM-LH21486B. H. MCCM-LH21486B, shape indicated by arrows. I. MCCM-LH21486B, broken bone showing slight pitting. J. MCCM-LH21486B, bone with corrosion lines. K. Ellipsoidal coprolite, MCCM-LH15929b. L. MCCM-LH15929b, lamellated bone structure indicated by arrows. M. MCCM-LH15929b, putative bone showing tubercles as ornamentation of a ?ganoid scale, indicated by arrows. N. Elongated coprolite, MCCM-LH-LI15-001B. O. MCCM-LH-LI15-001B, bone showing noticeable pitting, indicated by arrows.


Fig. 3. SEM and BSE images of coprolite specimens from the Las Hoyas locality (upper Barremian, Cuenca, Spain). A. Fir-tree coprolite, MCCM-LH32378 B. Putative tooth, MCCM-LH32378. C. Irregular coprolite, MCCM-LH28253a. D. Ostracod valve, MCCM-LH28253a. E. Broken putative vertebrae indicated by arrows, MCCM-LH28253a. F. Rosary coprolite, MCCM-LH23541A. G. Putative broken ribs, MCCM-LH23541A. H. Straight lace coprolite, MCCM-LH20198b. I. Big vertebra with lamellar bone, MCCM-LH20198b. J. Coali-fied plant structure, MCCM-LH20198b. K. Thin lace coprolite, MCCM-LH-GQ17-030a. L. Striated bone, MCCM-LH-GQ17-030a. M. Articulated vertebrae indicated by arrows, MCCM-LH-GQ17-030a. N. Overlapping bones, MCCM-LH-GQ17-030a. O. Thin lace coprolite, MCCM-LH-LI15-028B. P. Vertebrae with broken ribs anchored, MCCM-LH-LI15-028B. Q. Probable fish palate indicated by arrows, MCCM-LH-LI15-028B.


Mor

phot

ype

Spec

imen

num

ber

Cop

rolit

e si

ze

(mm

)N

IIS

(mm

)ID

IAPC

FEC

S

Bum

p-he

aded

lace

MC

CM

-LH

1599

3a(F

ig. 2

A–D

)

Leng

th: 3

9.6

Dia

met

er: 3

.3

mid

par

t; 9

bulg

e

>45

0.2–

0.9

Nat

ure:

Ver

tebr

ate

(fish

). El

emen

ts:

plan

t par

ts: S

ectio

n of

a h

ollo

w

verte

bra

and

para

llel fi

bres

, veg

etal

ep

ider

mis

Frag

men

ted

and

over

lapp

edPi

tting

No

corr

osio

n lin

esA

ll ty

pes

Ligh

t

Bum

p-he

aded

lace

MC

CM

-LH

3539

3Le

ngth

: 40

Dia

met

er: 3

mid

pa

rt; 9

bul

ge

>45

0.4–

0.8

In th

e bu

lge:

1.3

–1.6

Nat

ure:

Ver

tebr

ate

(fish

): br

anch

ial

arch

, rib

sFr

agm

ente

d an

d ov

erla

pped

Pitti

ngSo

me

corr

osio

n lin

es

Irre

gula

r and

st

raig

htLi

ght

Circ

ular

M

CC

M-L

H21

425b

Dia

met

erca

. 4.4

<15

0.2–

0.9

Larg

est i

nclu

sion

: 1.

6

Nat

ure:

Bon

y fr

agm

ents

Isol

ated

Pitti

ngN

o co

rros

ion

lines

All

type

sLi

ght

Circ

ular

M

CC

M-L

H26

943b

Dia

met

erca

. 5.5

31–4

50.

3–1.

1N

atur

e: V

erte

brat

e (fi

sh):

Fish

scal

esIs

olat

edPi

tting

Som

e co

rros

ion

lines

All

type

sLi

ght

Cyl

inde

r M

CC

M-L

H94

75A

(Fig

. 2E–

F)

Leng

th: 4

9D

iam

eter

: 14.

5<1

5N

ot a

t mac

rosc

ale

Nat

ure:

Ver

tebr

ate

(?fis

h); a

rthro

pod,

m

ollu

sc (g

astro

pod)

part=

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ated

: seq

uenc

e of

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eth

inde

term

inat

e or

the

inte

rnal

mou

ld o

f a

tiny

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pod

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app

licab

leN

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pplic

able

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r M

CC

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(Fig

. 2G

–J)

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th: 5

0.3

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met

er: 9

31–4

51.

2–4.

6La

rges

t inc

lusi

on:

5.7

Nat

ure:

Ver

tebr

ate

(fish

): Fi

sh sc

ales

Isol

ated

Pitti

ngC

orro

sion

line

sA

ll ty

pes

Mod

erat

e(p

ittin

g m

ore

exte

nsiv

e th

an

corr

osio

n)C

ylin

der

MC

CM

-LH

2871

9aLe

ngth

: 41.

2D

iam

eter

: 11

<15

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at m

acro

scal

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e: S

mal

l and

isol

ated

bon

y in

clus

ion

Not

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Not

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licab

leN

ot a

pplic

able

Cyl

inde

r M

CC

M-L

H-L

I15-

019

Leng

th: 1

4D

iam

eter

: 3.8

>45

0.2–

1.1

Nat

ure:

Ver

tebr

ate

(fish

): Po

ssib

le fi

sh

ribs a

nd a

fish

scal

e in

sect

ion

cut

Isol

ated

Pitti

ngN

o co

rros

ion

lines

All

type

sLi

ght

Con

eM

CC

M-L

H16

609B

Leng

th: 1

5.5

Dia

met

er: 9

15–3

00.

2–2.

2N

atur

e: V

erte

brat

e (fi

sh):

Scal

es a

nd

bony

rem

ains

Isol

ated

Pitti

ngC

orro

sion

line

sB

lunt

and

st

raig

htM

oder

ate

(cor

rosi

on

grea

ter t

han

pitti

ng)Ta

ble

2

Cha

ract

eriz

atio

n of

the

Las H

oyas

cop

rolit

e as

soci

atio

n: si

ze, i

nclu

sion

feat

ures

, and

pat

tern

s of t

he d

iges

tive

chem

ical

pro

cess

es.

Abb

revi

atio

ns: N

I – n

umbe

r of i

nclu

sion

s; IS

– in

clus

ion

size

; ID

– in

clus

ion

iden

tifica

tion;

IA –

incl

usio

n ar

rang

emen

t;

PC –

pitt

ing

and

corr

osio

n; F

E –

shap

e of

the

frac

ture

s obs

erve

d at

the

ends

; CS

– ch

emic

al si

gnal

.


Mor

phot

ype

Spec

imen

num

ber

Cop

rolit

e si

ze

(mm

)N

IIS

(mm

)ID

IAPC

FEC

S

Ellip

soid

al

MC

CM

-LH

1592

9b(F

ig. 2

K–M

)

Leng

th: 1

0D

iam

eter

: 430

–45

0.5–

1.7

Larg

est i

nclu

sion

: 6.

8

Nat

ure:

Ver

tebr

ate

(fish

): ?g

anoi

d sc

ale,

oth

ers w

ith la

mel

late

d bo

ne

stru

ctur

e

Isol

ated

and

frag

-m

ente

dPi

tting

Som

e co

rros

ion

lines

All

type

sLi

ght

Elon

gate

d M

CC

M-L

H-L

I15-

001B

(Fig

. 2N

–O)

Leng

th: 1

4.2

Dia

met

er: 6

.615

–30

0.7–

2.9

Nat

ure:

Bon

y fr

agm

ents

Isol

ated

Pitti

ngC

orro

sion

line

sB

lunt

and

pi

nche

dM

oder

ate

(pitt

ing

grea

ter

than

cor

rosi

on)

Elon

gate

d M

CC

M-L

H-L

I15-

002A

Leng

th: 1

3.2

Dia

met

er: 8

.315

–30

0.3–

4.3

Nat

ure:

Bon

y fr

agm

ents

Isol

ated

Pitti

ngC

orro

sion

line

sB

lunt

Mod

erat

e(p

ittin

g gr

eate

r th

an c

orro

sion

)Fi

r-tre

e M

CC

M-L

H32

378B

(Fig

. 3A

–B)

Leng

th: 9

Dia

met

er: 4

.2>4

50.

2–1

Nat

ure:

Ver

tebr

ate:

Sm

all t

ooth

and

bo

ny fr

agm

ents

Isol

ated

and

frag

-m

ente

dPi

tting

Som

e co

rros

ion

lines

All

type

sLi

ght

Irre

gula

r M

CC

M-L

H20

124

Leng

th: 4

0D

iam

eter

: 26.

5<1

5N

ot a

t mac

rosc

ale

Nat

ure:

Sm

all i

sola

ted

tubu

lar i

nclu

-si

onN

ot a

pplic

able

Not

app

licab

leN

ot a

pplic

able

Not

app

licab

le

Irre

gula

rM

CC

M-L

H27

015A

Leng

th: 3

0D

iam

eter

: 12.

515

–30

0.5–

13N

atur

e: B

ony

frag

men

ts

Isol

ated

and

frag

-m

ente

dG

reat

pitt

ing

Cor

rosi

on li

nes

Blu

ntH

eavy

Irre

gula

r M

CC

M-L

H28

253A

(Fig

. 3C

–E)

Leng

th: 1

5.5

Dia

met

er: 4

.2>4

50.

2–0.

6N

atur

e: V

erte

brat

e (fi

sh);

Elem

ents

: C

rust

acea

n (o

stra

cod,

?C

ypri

dea)

: ve

rtebr

ae a

nd ri

bs; v

alve

Frag

men

ted

Slig

ht p

ittin

gN

o co

rros

ion

lines

All

type

sLi

ght

Ros

ary

MC

CM

-LH

2354

1A(F

ig. 3

F–G

)

Leng

th: 2

3D

iam

eter

: 5.3

>45

0.4–

2.2

Nat

ure:

Ver

tebr

ate

(?fis

h): B

roke

n rib

s Fr

agm

ente

dN

o pi

tting

Som

e co

rros

ion

lines

Irre

gula

rLi

ght

Spira

l M

CC

M-L

H22

349

Leng

th: 1

0.4

Dia

met

er: 4

.8<1

50.

3 –0.

5N

atur

e: T

wo

smal

l inc

lusi

ons

Isol

ated

Not

app

licab

leN

ot a

pplic

a-bl

eN

ot a

pplic

a-bl

eSt

raig

ht la

ce

MC

CM

-LH

1661

1ALe

ngth

: 11.

5D

iam

eter

: 2>4

50.

2 –0.

8N

atur

e: V

erte

brat

epa

rt: b

roke

n ?v

erte

bra

Frag

men

ted

Slig

ht p

ittin

gSo

me

corr

osio

n lin

es

All

type

sLi

ght

Stra

ight

lace

MC

CM

-LH

2019

8B(F

ig. 3

H–J

)

Leng

th: 2

8D

iam

eter

: 2.8

>45

0.2–

0.8

Nat

ure:

Ver

tebr

ate

(fish

); El

e-m

ents

: pla

nt p

arts

: fish

ver

tebr

a w

ith la

mel

lar b

one,

coa

lified

pla

nt

stru

ctur

e, tu

bula

r bon

y st

ruct

ures

(p

roba

bly

ribs)

Frag

men

ted

Slig

ht p

ittin

gN

o co

rros

ion

lines

All

type

sLi

ght

Thin

lace

M

CC

M-L

H16

564A

Leng

th: 1

9.5

Dia

met

er: 2

>45

0.3 –

1.2

Nat

ure:

Bon

y fr

agm

ents

Frag

men

ted

Slig

ht p

ittin

gN

o co

rros

ion

lines

All

type

sLi

ght


Mor

phot

ype

Spec

imen

num

ber

Cop

rolit

e si

ze

(mm

)N

IIS

(mm

)ID

IAPC

FEC

S

Thin

lace

M

CC

M-L

H-G

Q-

17-0

30A

(Fig

. 3K

–N)

Leng

th: 1

00D

iam

eter

: 2.2

>45

0.3–

1.3

Nat

ure:

Ver

tebr

ate

(fish

): Se

vera

l gr

oups

of a

rticu

late

d ve

rtebr

ae,

fish

rays

and

ribs

Frag

men

ted,

ov

erla

pped

and

ar

ticul

ated

Pitti

ngSo

me

corr

osio

n lin

es

All

type

sLi

ght

Thin

lace

M

CC

M-L

H-L

I15-

028B

(Fig

. 3O

–Q)

Leng

th: 2

5D

iam

eter

: 2.5

>45

0.2–

0.7

Nat

ure:

Ver

tebr

ate

(fish

): Ve

rte-

brae

(som

e pa

rtial

ly a

rticu

late

d w

ith a

ncho

red

ribs)

, rad

ius f

rom

fin

s and

a p

alat

e fr

om m

acro

sem

ii-di

d fis

h (?

Not

agog

us) o

r a ju

veni

le

tele

ost

Frag

men

ted

No

pitti

ngSo

me

corr

osio

n lin

es

All

type

sLi

ght

Tabl

e 2

Con

tinua

tion

from

the

form

er p

ages

.

Fig. 4. Box-plot graphic representing size values of inclusions for the different morphotypes. Circles represent the outliers for the corresponding morphotype, the box represents the percentiles, the thin line represents the median, and whiskers represent the minimal and maximal values.

Non-Metric Multidimensional Scaling (NMDS) analysis

The NMDS analysis clearly separates the morphotypes into at least three groups. The NMDS has a stress value of 0.085 and is therefore reliable for describing the relation-ships among the samples in two-dimensional ordination plots (Clarke and Warwick, 2001; Zuschin et al., 2006). In Figure 6, the dots that are close together represent mor-photypes that are very similar, regarding the taphonomic properties of the inclusions; the dots that are far apart have different values and even in the sample some have missing values (i.e., 18SPI, 7 and 5 CYL and 14 IRR in Fig. 6). The NMDS shows the interplay between the number of in-clusions and their arrangement in the coprolite matrix along the y-axis, and the intensity of chemical signals along the x-axis of the graph. The group of positive y values and neg-ative x values defines coprolites that are densely filled and have fragmented to overlapped inclusions, with light chem-ical signals (bump-headed lace (BHL); fir-tree (FIR); rosary (RO); straight lace (STR); and thin lace (TH) morphotypes). The second group (circular (CIR); spiral (SPI); and cylinder (CYL) morphotypes) are scarcely filled, with isolated in-clusions, and slight chemical signals. The third group, com-prising the cone (CO); and elongated (ELO) morphotypes, occupies the positive x values, because its members share a moderate degree of corrosion and pitting.

A minimum spanning tree was also constructed to show the total minimal distance connecting all points, to visualize the grouping close points. All the same morphotypes, ex-cept for irregular (IRR), are placed within the same group. Closer points are in the upper left of the plot, corresponding also to coprolites with lower diameters (Fig. 5); note that the coprolite diameter increases towards the positive values of the x-axis (see Figs 5, 6).


Fig. 5. Graphic showing the relationship between size values of inclusions and coprolite maximum diameter. The data are given rep-resenting every morphotype by means of different colours. For bump-headed lace coprolites, the diameter represented is the diameter of the coprolite cord.

DISCUSSION

The present study was designed to establish the type of correlation that exists between the coprolite morphotypes and the alterations in the coprolite inclusions, due to the di-gestive strategies of the producers. In order to arrange a rep-resentative sample (e.g., Rodríguez-de la Rosa et al., 1998; Luo et al., 2017; Dentzien-Diaz et al., 2018) and to test the similarity of the observed taphonomic properties of the inclusions, more than one specimen for each morphotype was included (except for cone, ellipsoidal, fir-tree, rosary and spiral morphotypes) and around 500 inclusions were counted and observed, combining lower magnifications and SEM, to provide a more relevant taphonomic description (Butler and Schroeder, 1998).

The features observed in the specimens show enough shared similarities to make some generalizations about the morphotypes and discuss how these are ordered. This is particularly relevant for the circular, cylinder, bump-head-ed lace, elongated, straight lace and thin lace morphotypes. In the irregular morphotype, the variation of the taphonom-ic features and the type of inclusions were not consistent among specimens. This variation indicates that the morpho-type could have been a compilation of non-definable shapes, ingestions and/or producers. In these cases and in the ones represented by only one specimen (cone, ellipsoidal, fir-tree, rosary and spiral), new specimens should be analysed to confirm the degree of consistency of digestive alteration among specimens per morphotype.

The sizes of the Las Hoyas coprolites and the fact that they mostly contain bone fragments suggest that most were

Fig. 6. Non-metric Multidimensional Scaling (NMDS) ordi-nation. The graph represents the relationships among samples ac-cording to the taphonomic properties analyzed for the coprolite inclusions. The line connecting the dots is the Minimum Span-ning Tree. DS1 = digestive strategy 1; DS2 = digestive strategy 2; DS3 = digestive strategy 3.


produced by vertebrate predators (Barrios-de Pedro et al., 2018a, b). All the selected taphonomic properties were com-bined to assess the ways of ingestion and the digestive pro-cesses, and to furnish the different digestive strategies found from the coprolite association.

Way of ingestion

The term ‘way of ingestion’ refers to the mechanism used to ingest the prey (e.g., swallowing, chewing, crushing, tear-ing apart, chopping, cutting; e.g., Diefenbach, 1975; Steven and Hume, 1995; Gutiérrez, 1998; Fernández-Jalvo and An-drews, 2016). The producers of the coprolite morphotypes, showing a higher number of inclusions (> 45) and with partially fragmented and occasionally overlapped remains, could have ingested their food with limited mechanical pro-cessing of the food in the mouth. In the thin lace morpho-type, some of the inclusions even appear to be articulated in successive packages (Fig. 7), indicating that the prey would have been swallowed whole. In contrast, the producers of coprolites with relatively low numbers of inclusions (< 45) and with mostly unidentifiable remains probably would have masticated (chewed, torn apart, chopped or cut) their prey. The way of ingestion seems to be relevant because food breakage increases the effects of the digestion process, allowing digestive juices to penetrate into cavities, and then increasing the bone area exposed to gastric acids (He and Wurtsbaugh, 1993; Fernández-Jalvo and Andrews, 2016).

Digestive process

The term ‘digestive process’ refers to the chemical signals left during digestion. The levels of acidity in the predators’ stomachs and the type of digestive enzymes are two fac-tors controlling digestion. The former is the main modifying agent of the surfaces of inclusions and requires time to oper-ate, while the latter breaks down the organic constituents of bones, modifying their internal structure (e.g., Diefenbach, 1975; Butler and Schroeder, 1998; Pinto-Lona and Andrews, 1999; Fernández-Jalvo and Andrews, 2016). In general, the inclusions are more easily identified in morphotypes that show a minor degree of chemical signals (bump-headed lace, ellipsoidal, fir-tree, rosary, straight lace and thin lace), while in those exhibiting a moderate degree of chemical signals the identification of inclusions is difficult or almost impossible. The most identifiable elements are scales and vertebrae, but it was impossible to provide a list of bony elements by skeletal part (see Butler and Schroeder, 1998), even for the cases with a higher abundance of inclusions, because they are fragmented, deformed, and compressed.

The modifications due to digestion (e.g., the shape of the edges of the inclusions, and the splitting and thinness of the bone surface) are the most valuable evidence of diet (Andrews and Fernández-Jalvo, 1998; Pinto-Lona and An-drews, 1999). The features have been mostly applied in the study of archaeological sites (e.g., Pinto-Lona and Andrews, 1999; Tappen and Wrangham, 2000). The present authors

Fig. 7. Detail of the central part of the thin lace coprolite MCCM-GQ17-030A, showing packages of inclusions (dashed lines) with articulated elements (indicated by arrows). Image taken with a 3D high-resolution microscope with interferometric and profilometry in a focus variation mode.

The morphotypes rich in undigested food remains (bump-headed lace, fir-tree, rosary, straight lace, and thin lace) display a similar size range of inclusions and the mean value of the inclusions is the smallest (around 1 mm; Fig. 4). This, together with the observation that the inclu-sions identified correspond mostly to small fishes, indicates that the producers probably fed on the same sort of prey. In the scarcely filled morphotypes (cylinder, cone, ellipsoi-dal, and elongated), the range of the size of inclusions is heterogeneous and the larger inclusions even reach more than 4 mm in length. The producers of such morphotypes probably would have fed on a different kind of prey, most likely slightly larger animals than those in the first group of morphotypes (Fig. 5), which would indicate a relationship between the size of the prey and the coprolite producer.

have evaluated some of these features, but they are obvious-ly more difficult to discern since the inclusions are partially covered and some even overlap. Nonetheless, in general, the Las Hoyas coprolite association is characterized by bearing a minor degree of corrosion and pitting, and most of the fractures at the ends of inclusions are varied and even in-clude morphotypes with sharp or rectilinear edges (Fig. 3L). The strongest action of acids, resulting in round and smooth edges of inclusions, the splitting of bone surfaces with lon-gitudinal lines following the bone weakness, and the thin-ning of bone walls, were observed in the irregular, cone and elongated morphotypes. Thinness leads to inward curving and the collapsing of the thinned edges of bones (Pinto-Lo-na and Andrews, 1999), a condition observed, for instance, in specimen MCCM-LH28253a (Fig. 3E).


Digestive strategies

NMDS analysis (Fig. 6) provided the basis for outlining the different digestive strategies of the Las Hoyas ecosys-tem with regard to the coprolite association. The diges-tive strategies can be distinguished by linking food intake and digestion process. The first one (Fig. 6, DS1) includes a diverse number of morphotypes and owing to the com-pleteness and abundance of inclusions the strategy would correspond to animals with low digestive assimilation. It is characterized by a food intake of one or several prey with limited processing in the mouth. The food eaten would be accumulated in the digestive system for a short time and/or the digestive process would exhibit low acid content (Owocki et al., 2012; Luo Mao et al., 2017). This strate-gy would only make sense in a scenario, where food was very abundant and nourishment was guaranteed (Cork and Kenagy, 1989). Of particular interest is the thin lace copro-lite MCCM-LH-GQ17-030A (22TH in Fig. 6), because the producer ingested more than one prey in a short period of time, which indicates the special voracity of the predator, compared with other producers of the same coprolite mor-photype.

The second digestive strategy (Fig. 6, DS2) contains a limited number of morphotypes, and owing to the reduc-tion and breakage of undigested food, the strategy indicates an improved digestive assimilation. It is characterized by mastication prior to deglutition that might be enhanced by gastric motility in the stomach (Diefenbach, 1975; Daven-port et al., 1990). It also would be associated with a longer retention time and food processing by the digestive system of the producer, which could indicate a more effective and/or longer digestion (Steven and Hume, 1995; Luo Mao et al., 2017). A longer digestion would be enhanced by in-creasing the volume and the length of the digestive system of the predators. This fact would favour the retention time of food in the organism and therefore increase the diges-tion rate and the absorption of nutrients in the intestine (Bozinovic, 1993).

A possible third digestive strategy (Fig. 6, DS3) has a limited number of morphotypes, and the fragmented inclu-sions with abundant chemical signals indicate an efficient digestive strategy. It combines scarcity of food remains, large inclusions in coprolites with wider diameters, and im-portant chemical signals of pitting and corrosion and even the blunting of edges. This combination indicates that the mechanism used to ingest the prey could have also included mastication (e.g., crushing and tearing apart), but above all a more efficient chemical digestion with food retained for a long time in the digestive system.

Feeding groups

The feeding ecology of the Las Hoyas Barremian ecosys-tem, on the basis of the coprolite association, can be charac-terized as: 1) a lentic environment enriched in prey, owing to the abundance of inclusions, as is reflected in the body fossil record; 2) having a greater diversity in morphotypes with a swallowing or non-processed ingestion; 3) having a limited diversity of morphotypes, greater in diameter, with varied

mastication ingestion and rather more efficient digestion. The swallower group contains a variety of thin-narrow and long coprolites that could have been produced by species of animals able to swallow whole prey such as fishes, frogs, birds, and lizards. The other group would include larg-er predators able to capture and process larger prey. From all the potential producers, it is clear that crocodiles could have formed part of this group. They ingest food in large pieces, with minor mechanical breakdown in the mouth; the reduction in size of the food and its chemical degradation occurs mainly in the stomach, enhanced by the motility of this organ and by gastroliths; and they retain the food in the digestive system for a long period of time (Diefenbach, 1975; Davenport et al., 1990). Morphotypes included in the third strategy (elongated to conical) match the shape of ex-tant crocodilian faeces (Milán, 2012).

CONCLUSIONSThe study of inclusions in coprolites provides informa-

tion not only about the prey that the producers ingested, but also about the digestive processes of the predators. The study of digestive alterations previously was applied mainly in coprolites found at archaeological sites. In this study, the authors have applied non-destructive techniques to identify, quantify, and estimate the degree of alteration of coprolite inclusions, which reflects the intake and the diges-tive secretions of the coprolite producers. This has allowed characterization of the whole Las Hoyas coprolite associa-tion, the producers of which were predominately vertebrate predators. Although the authors have found some ostracod and plant remains, they have not been able to verify the presence of insects or crustaceans in the coprolites. Given the great diversity of coprolite morphotypes found in this lacustrine environment, this study has allowed the estab-lishment of correspondence between morphotypes and the types of alteration of the inclusions due to digestion. The results show that there is no unambiguous correspondence between morphotypes and inclusion properties, as there are several morphotypes that share the same type of digestive features in their inclusions. The features used have been useful in ordering and characterizing different types of di-gestive strategies. Accordingly, the authors propose up to three types of digestive strategy for the coprolite produc-ers: the first involves a limited processing in the mouth, a short retention time of food, and/or a low efficiency of the digestive system; the second implies possible mastication of food prior to deglutition, followed by a longer retention time of food in the digestive system; and the third includes a group with mastication, long retention time of food, and more effective digestion. This contribution is a first step in the understanding of the feeding ecology of this Barremian wetland ecosystem, based on the coprolite association.

Acknowledgements

The authors thank Santiago Langreo and Mercedes Llandres at the Museo de las Ciencias de Castilla-La Mancha for allowing ac-cess to the coprolite collection. Special thanks go to Alberto Jorge and Marta Furio at the Museo Nacional de Ciencias Naturales for


carrying out the SEM analyses; as well as to Cristina Paradela for carrying out the 3D surface measurement with the 3D high-resolu-tion microscope. Thanks also go to Miguel Angel Fernandez of the Department of Biology at the Universidad Autónoma de Madrid for the digital camera. Comments and correction of the original manuscript by Spencer G. Lucas (New Mexico Museum of Nat-ural History and Sciences) and Martin Qvarnström (Uppsala Uni-versity, Sweden) improved the study. Funds were provided by the MINECO reference CGL2013-42643-P. The fellowship reference BES-2014-070985 of the Program for the Training of Researchers of the MINECO is associated with this project.

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Specimen NI PT CL CS DM AI FE1BHL 4 2 1 1 1 2 52BHL 4 2 2 1 1 2 33CIR 1 2 1 1 1 4 54CIR 3 2 2 1 2 4 55CYL 1 ? ? ? 3 1 ?6CYL 3 2 2 2 2 4 57CYL 1 ? ? ? 3 1 ?8CYL 4 2 1 1 1 4 59CON 2 2 2 2 2 4 110ELL 3 2 2 1 1 2 511ELO 2 3 2 2 2 4 112ELO 2 3 2 2 2 4 113FIR 4 2 2 1 1 2 514IRR 1 ? ? ? 4 1 ?15IRR 2 4 2 3 3 2 116IRR 4 2 1 1 1 2 517ROS 4 1 2 1 2 2 418SPI 1 ? ? ? 1 4 ?19STR 4 2 2 1 1 2 520STR 4 2 1 1 1 2 521TH 4 2 1 1 1 2 522TH 4 2 2 1 1 2 523TH 4 1 2 1 1 2 5

Appendix 1

Values of the features used for the non-metric multidimensional scaling (NMDS) analysis. BHL – bump-headed lace; CIR – circular; CYL – cylinder; CON – cone; ELL – ellipsoidal; ELO – elongated;

FIR – fir-tree; IRR – irregular; ROS – rosary; SPI – spiral; STR – straight lace; TH – thin lace; NI – number of inclusions (1 <15 ; 2 = 15–30 ; 3 = 31 – 45; 4 >45); PT – pitting (1 – absent; 2 – light;

3 – noticeable; 4 – great); CL–corrosion lines (1 – absent; 2 – slight; 3 – noticeable); CS – chemical signal (1 – light; 2 – moderate; 3 – heavy); DM – diameter in mm (1 < 5; 2 = 5–9.9 ; 3 = 10–19.9; 4 >20); AI – inclusion arrangement

(1 – no visible; 2 – fragmented; 3 – articulated; 4 – isolated); FE – shape of the fractures observed at the ends (1 – blunt; 2 – pinched; 3 – straight; 4 – irregular; 5 – all types).


Bump-headed lace coproliteso MCCM-LH15993a (Fig. 2A–D)• Number of inclusions: >45.• Inclusions and coprolite size: Coprolite length is 39.6

mm. Coprolite diameter is about 3.3 mm in the body and 9 mm at the bulge. Inclusions range from 0.2 to 0.9 mm.

• Identification of inclusions: Identified inclusions correspond to the section of a hollow fish vertebra, a plant fragment with parallel fibres, and a possible plant epidermis with a cellular size of about 20 microns. Most of the remains appear fragmented and overlapping.

• Action of digestive process: Inclusions are partially al-tered with slight pitting. Corrosion lines are absent, and fractures at the ends of inclusions are of all types. The degree of corrosion is light.

o MCCM-LH35393• Number of inclusions: >45.• Inclusions and coprolite size: Coprolite length is

40 mm. Coprolite diameter is around 3 mm in the body and 9 mm in the bulge. Inclusions usually range from 0.4 to 0.8 mm. Largest inclusions at the bulging end measure 1.3–1.6 mm.

• Identification of inclusions: Identified inclusions corre-spond to a branchial arch and possible broken fish ribs. Most of the remains appear fragmented and overlapping; occasionally the bigger inclusions appear isolated.

• Action of digestive process: Inclusions are partially altered with slight pitting. Some of the bigger bones present corrosion lines. Fractures at the ends of inclu-sions are mostly irregular or straight. Some remains are broken and chipped. The degree of corrosion is light.

Circular coproliteso MCCM-LH21425b • Number of inclusions: <15.• Inclusions and coprolite size: Coprolite length/diame-

ter is around 4.4 mm. Inclusions range from 0.2 to 0.9 mm. The largest inclusion is around 1.6 mm.

• Identification of inclusions: All of them are bony inclu-sions. The remains appear isolated.

• Action of digestive process: Inclusions are partially altered with slight pitting. Corrosion lines are absent and fractures at the ends of inclusions are of all types. The degree of corrosion is light.

o MCCM-LH26943b • Number of inclusions: 31–45.• Inclusions and coprolite size: Coprolite length/diame-

ter is around 5.5 mm. Inclusions range from 0.3 to 1.1 mm.

• Identification of inclusions: Identified inclusions correspond to scales. Most of the remains appear isolated.

Appendix 2

Detailed description of inclusion characteristics.

• Action of digestive process: Inclusions are partially altered with slight pitting and some of them exhibit corrosion lines as well. Fractures at the ends of inclusions are of all types. The degree of corrosion is light.

Cylinder coproliteso MCCM-LH9475A (Fig. 2E–F)• Number of inclusions: N <15.• Inclusions and coprolite size: Coprolite length is

49 mm. Coprolite maximum diameter is 14.5 mm.• Identification of inclusions: This specimen apparently

does not exhibit inclusions on its surface. A small isolat-ed inclusion was identified as being like a sequence of teeth or the internal mould of a tiny, grooved, unde-termined fragment, maybe a snail or an arthropod.

• Action of digestive process: Not applicable. • Plate ‘b’ is not preserved.

o MCCM-LH21486B (Fig. 2G–J)• Number of inclusions: 31–45.• Inclusions and coprolite size: Coprolite length is

0.3 mm. Coprolite maximum diameter is 9 mm. Inclu-sions range from 1.2 to 4.6 mm. A larger inclusion in the middle of the coprolite matrix measures 5.7 mm.

• Identification of inclusions: Four inclusions have been identified as fish scales because of their fusiform shape. The other bony inclusions are isolated in the coprolite matrix.

• Action of digestive process: Fish scales show slight pitting and no corrosion lines; they are probably section cuts of scales. The biggest inclusion presents slight pit-ting and corrosion lines, with the biggest lines filled with a mineral (probably calcite). The most elongated inclusions show slight pitting over the entire surface and lots of corrosion lines. Fractures at the ends of inclusions are of all types. The degree of corrosion is moderate.

o MCCM-LH28719A • Number of inclusions: N <15.• Inclusions and coprolite size: Coprolite length is

41.2 mm. Coprolite maximum diameter is 11 mm.• Identification of inclusions: This specimen apparently

does not exhibit inclusions on its surface. In SEM a small and isolated inclusion, perhaps representing bone, was observed.

• Action of digestive process: Not applicable. • This specimen is an exception to the common way of

naming the fossils. It has a letter at the end of its speci-men number, being an isolated coprolite with its dorsal surface exposed.


o MCCM-LH-LI15-019• Number of inclusions: >45.• Inclusions and coprolite size: Coprolite length is

14 mm. Coprolite maximum diameter is 3.8 mm. Inclu-sions range from 0.2 to 1.1 mm.

• Identification of inclusions: Identified inclusions are tubular inclusions (possibly fish ribs) and another one that could be a fish scale in cross-section. Inclusions appear isolated.

• Action of digestive process: Some inclusions exhibit slight pitting on their surfaces and an absence of corro-sion lines. Fractures at the ends of the inclusions are of all types. The degree of corrosion is light.

Cone coproliteso MCCM-LH16609B• Number of inclusions: 15–30.• Inclusions and coprolite size: Coprolite length is

15.5 mm. Coprolite maximum diameter is around 9 mm. There is a wide inclusion range, from 0.2 to 2.2 mm.

• Identification of inclusions: The only identified inclu-sions are fragments of fish scales and bony pieces. Inclusions appear isolated.

• Action of digestive process: The bony tubular inclu-sions and the fish scales show slight pitting. Bones present corrosion lines on their surface. The deeper lines were produced probably during the rock splitting into part and counterpart. Fractures at the ends of the inclu-sions are blunt and straight; most of them are covered by coprolite matrix. The degree of corrosion is moderate.

Ellipsoidal coproliteso MCCM-LH15929b (Fig. 2K–M)• Number of inclusions: 30–45.• Inclusions and coprolite size: Coprolite length is

10 mm. Coprolite maximum diameter is 4 mm. Inclusions range from 0.5 to 1.7 mm. The largest inclusion at the edge of the matrix measures 6.8 mm.

• Identification of inclusions: Bony inclusions appear both isolated and fragmented. One of the inclusions shows tubercles as ornamentation of a ?ganoid scale, while others exhibit a lamellated bone structure.

• Action of digestive process: Most of the inclusions show both slight pitting and slight corrosion lines on their surface. Fractures at the ends of the inclusions are of all types. The degree of corrosion is light.

Elongated coproliteso MCCM-LH-LI15-001B (Fig. 2N–O)• Number of inclusions: 15–30.• Inclusions and coprolite size: Coprolite length is

14.2 mm. Coprolite maximum diameter is 6.6 mm. Inclusions range from 0.7 to 2.9 mm.

• Identification of inclusions: Inclusions are bony, isolated fragments, perpendicular to the coprolite matrix.

• Action of digestive process: Inclusions present notice-able pitting on their surface, with presence of corrosion lines. Fractures at the ends of the inclusions are both blunt and pinched. The degree of corrosion is moderate.

o MCCM-LH-LI15-002A• Number of inclusions: 15–30.• Inclusions and coprolite size: Coprolite length is

13.2 mm. Coprolite maximum diameter is about 8.3 mm. There is a wide inclusion range that varies from 0.3 to 4.3 mm.

• Identification of inclusions: Inclusions are unidentifi-able bony remains. Most of them appear isolated.

• Action of digestive process: Inclusions present notice-able pitting on their surface, with the presence of slight corrosion lines. Fractures at the ends of the inclusions are mostly blunt. The degree of corrosion is moderate.

Fir-tree coproliteso MCCM-LH32378B (Fig. 3A–B)• Number of inclusions: >45.• Inclusions and coprolite size: Coprolite length is about

9 mm. Coprolite maximum diameter is 4.2 mm. Inclu-sions range from 0.2 to 1 mm.

• Identification of inclusions: The only identified inclu-sion corresponds to a tooth. The remains appear both isolated and fragmented.

• Action of digestive process: Inclusions are partially altered with slight pitting, and some of them exhibit cor-rosion lines as well. Fractures at the ends of inclusions are of all types. The degree of corrosion is light.

Irregular coproliteso MCCM-LH20124• Number of inclusions: N <15.• Inclusions and coprolite size: Coprolite length is

40 mm. Coprolite maximum diameter is about 26.5 mm.• Identification of inclusions: This specimen apparently

does not exhibit inclusions at its surface. At SEM a small and isolated tubular fragment was observed.

• Action of digestive process: Not applicable.

o MCCM-LH27015A• Number of inclusions: 15–30.• Inclusions and coprolite size: Coprolite length is

around 30 mm. Coprolite maximum diameter is around 12.5 mm. Inclusions range from 0.5 up to 13 mm.

• Identification of inclusions: Inclusions are of a bony nature and appear both isolated and fragmented.

• Action of digestive process: In longer inclusions, pit-ting is slight (sometimes absent), but in the wider bony remains deeper holes are observed (great pitting). They show corrosion lines, although perpendicular breakages could have been enhanced by splitting. Most of the ends of inclusions are partially buried in the coprolite matrix, but some blunt ends are observed. The degree of corro-sion is heavy.

o MCCM-LH28253A (Fig. 3C–E)• Number of inclusions: >45.• Inclusions and coprolite size: Coprolite length is

around 15.5 mm. Coprolite maximum diameter is 4.2 mm. Inclusions range from 0.2 to 0.6 mm.


• Identification of inclusions: Identified inclusions are an ostracod valve (?Cypridea) and possible, very thin fish vertebrae and ribs. Inclusions appear fragmented.

• Action of digestive process: Pitting and corrosion lines are absent. Fractures at the ends of the inclusions are of all types. The degree of corrosion is light.

Rosary coproliteso MCCM-LH23541A (Fig. 3F–G)• Number of inclusions: >45.• Inclusions and coprolite size: Coprolite length is

around 23 mm. Coprolite maximum diameter is around 5.3 mm. Inclusions range from 0.4 to 2.2 mm.

• Identification of inclusions: Identified inclusions are broken ribs, probably from a fish. Inclusions appear fragmented.

• Action of digestive process: There is no pitting on the inclusions, just very thin corrosion lines. Fractures at the ends of inclusions are irregular (some inclusions are buried in the coprolite matrix). The degree of corrosion is light.

Spiral coproliteso MCCM-LH22349 • Number of inclusions: <15.• Inclusions and coprolite size: Coprolite length is

10.4 mm. Coprolite maximum diameter is 4.8 mm. The only two visible inclusions measure 0.3 and 0.5 mm.

• Identification of inclusions: Inclusions appear isolated. They are broken and partially covered by the coprolite matrix

• Action of digestive process: Not applicable.

Straight lace coproliteso MCCM-LH16611A • Number of inclusions: >45.• Inclusions and coprolite size: Coprolite length is

around 11.5 mm. Coprolite maximum diameter is around 2 mm. Inclusions range from 0.2 to 0.8 mm.

• Identification of inclusions: There is a broken inclusion that could be a vertebra. Inclusions appear fragmented.

• Action of digestive process: Some inclusions show both slight pitting and corrosion lines. Fractures at the ends of inclusions are of all types. The degree of corrosion is light.

o MCCM-LH20198B (Fig. 3H–J)• Number of inclusions: >45.• Inclusions and coprolite size: Coprolite length is


• Identification of inclusions: Identified inclusions are a transverse section of a fish vertebra with lamellar bone

(wider than the vertebra found in coprolite MCCM-LH15993a), a coalified plant structure, as well as tubular, bony structures (probably ribs). Inclusions appear fragmented.

• Action of digestive process: Neither slight pitting nor corrosion lines are observed. Fractures at the ends of the inclusions are of all types. The degree of corro-sion is light.

• Plate ‘a’ is not preserved.

Thin lace coproliteso MCCM-LH16564A• Number of inclusions: >45.• Inclusions and coprolite size: Coprolite length is

19.5 mm. Coprolite maximum diameter is around 2 mm. Inclusions range from 0.3 to 1.2 mm.

• Identification of inclusions: There are no identifiable inclusions. They appear fragmented.

• Action of digestive process: Neither slight pitting nor corrosion lines are observed. Fractures at the ends of the inclusions are of all types. The degree of corrosion is light.

o MCCM-LH-GQ17-030A (Fig. 3K–N)• Number of inclusions: >45.• Inclusions and coprolite size: Coprolite length is


• Identification of inclusions: Identified inclusions are several groups of articulated vertebrae, fish rays and ribs. Most of the inclusions appear fragmented and overlapped, and several articulated.

• Action of digestive process: Inclusions exhibit both slight pitting and corrosion lines. Fractures at the ends of the inclusions are of all types (sometimes they are buried in the coprolite matrix or under other inclusions). The degree of corrosion is light.

• Plate ‘b’ is not preserved.

o MCCM-LH-LI15-028B (Fig. 3O–Q)• Number of inclusions: >45.• Inclusions and coprolite size: Coprolite length is

around 25 mm. Coprolite maximum diameter is around 2.5 mm. Inclusions size ranges from 0.2 to 0.7 mm.

• Identification of inclusions: Identified inclusions are vertebrae (some of them seem to be partially articulated and with anchored ribs), possible radius from fins and a palate, which indicates that the possible prey was a macrosemiidid fish (?Notagogus) or a juvenile teleost. Most of the inclusions appear fragmented.

• Action of digestive process: Inclusions do not show pitting; only some of them show slight corrosion lines. Fractures at the ends of inclusions are of all types. The degree of corrosion is light.

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